Spectrum of germline mutations in smokers and non-smokers in Brazilian non-small-cell lung cancer (NSCLC) patients.

Lung cancer (LC) is a leading cause of cancer-related mortality. Although smoking is the major risk factor, ~15% of all cases occur in never-smokers, suggesting that genetic factors play a role in LC predisposition. Indeed, germline mutations in the TP53 gene predispose to multiple cancer types, including LC. To date, few studies compared the somatic and germline mutational profiles of LC cases by smoking status, and none was reported in Brazilians. Whole-exome sequencing (WES) was performed on two pools (seven smokers and six non-smokers) of tumor-derived DNA using the Illumina HiSeq2000 platform. Files from pools were analyzed separately using Ingenuity®Variant AnalysisTM and Mendel,MD. Validation of all candidate variants was performed by Sanger sequencing. Subsequently, validated mutations were analyzed in germline DNA from the same patients and in ethnically matched controls. In addition, a single recurring Brazilian TP53 germline mutation (R337H) was genotyped in 45 non-small-cell lung cancer patients.Four novel germline variants in the ATAD2, AURKA, PTPRD and THBS1 genes were identified exclusively in smoker patients, and four germline missense variants in PLCD1, RAD52, CP and CDC6 genes were identified solely in non-smokers. There were 4/45 (8.9%) germline carriers of the R337H TP53 mutation. In conclusion, the recurring Brazilian TP53 mutation should be genotyped in all non-small-cell lung cancer in Brazil, regardless of smoking status. Distinct pathogenic mutations and novel sequence variants are detected in Brazilian non-small-cell lung cancer patients, by smoking status. The contribution of these sequence variants to LC pathogenesis remains to be further explored.

Spectrum of somatic EGFR, KRAS, BRAF, PTEN mutations and TTF-1 expression in Brazilian lung cancer patients.

Lung cancer is the leading global cause of cancer-related mortality. Inter-individual variability in treatment response and prognosis has been associated with genetic polymorphisms in specific genes: EGFR, KRAS, BRAF, PTEN and TTF-1. Somatic mutations in EGFR and KRAS genes are reported at rates of 15-40% in non-small cell lung cancer (NSCLC) in ethnically diverse populations. BRAF and PTEN are commonly mutated genes in various cancer types, including NSCLC, with PTEN mutations exerting an effect on the therapeutic response of EGFR/AKT/PI3K pathway inhibitors. TTF-1 is expressed in approximately 80% of lung adenocarcinomas and its positivity correlates with higher prevalence of EGFR mutation in this cancer type. To determine molecular markers for lung cancer in Brazilian patients, the rate of the predominant EGFR, KRAS, BRAF and PTEN mutations, as well as TTF-1 expression, was assessed in 88 Brazilian NSCLC patients. EGFR exon 19 deletions (del746-750) were detected in 3/88 (3·4%) patients. Activating KRAS mutations in codons 12 and 61 were noted in five (5·7%) and two (2·3%) patients, respectively. None of the common somatic mutations were detected in either the BRAF or PTEN genes. TTF-1 was overexpressed in 40·7% of squamous-cell carcinoma (SCC). Our findings add to a growing body of data that highlights the genetic heterogeneity of the abnormal EGFR pathway in lung cancer among ethnically diverse populations.

Histomorphometric evaluation of the Ki-67 proliferation rate and CD34 microvascular and D2-40 lymphovascular densities drives the pulmonary typical carcinoid outcome.

Ki-67 has shown promise as a prognostic factor in pulmonary carcinoids. In this study, we sought to validate the importance of Ki-67 and study the relationships between Ki-67 and other stromal biomarkers of vascular density. We examined Ki-67, CD34, and D2-40 in tumor tissues from 128 patients with surgically excised typical carcinoid of the lung. We used immunohistochemistry and morphometry to evaluate the amount of tumor staining for cellular proliferation (Ki-67), microvascular density (CD34-MVD), and D2-40 lymphovascular density. The main outcome was overall survival, considered as life expectancy until death from metastasis. Specimens from patients with central tumors showed high CD34-MVD (P = .01), which was also significantly associated with a compromised surgical margin, lymph node metastasis, and clinical stage Ib. Equally significant was high D2-40 lymphovascular density in central specimens with a compromised surgical margin and lymph node metastasis. A high Ki-67 proliferation rate was significantly associated with tumors from patients with clinical stage IIb, IIIa, and IV disease. Multivariate Cox model analysis demonstrated that tumor location and stage, surgical margin, tumor size, and N stage were significantly related to survival time (P < .05). Quantitative staining of the tumor for Ki-67 and CD34-MVD served as prognostic factors (P < .05), which were more relevant than the surgical and pathological stage. Ki-67 greater than 5% and CD34-MVD greater than 7% staining comprise a subset of patients with higher death hazard; this outcome may harbor evidence for further prospective studies of target therapy after surgical resection.

Association between CLOCK, PER3 and CCRN4L with non­small cell lung cancer in Brazilian patients.

Circadian rhythms comprise of daily oscillations in a variety of biological processes and are regulated by an endogenous clock. Disruption of these rhythms has been associated with cancer progression, and understanding natural oscillations in cellular growth control, tumor suppression and cancer treatment, may reveal how clock and clock­controlled genes are regulated in normal physiological functioning. To investigate the association between clock genes and non­small cell lung cancer (NSCLC), we genotyped three tag SNPs (rs938836, rs17050680, rs3805213) in the Nocturnin gene (CCRN4L), five SNPs (rs228727, rs228644, rs228729, rs707467, rs104620202) in the period 3 (PER3) gene and one SNP (rs6855837) in the CLOCK gene, in 78 Brazilian patients with NSCLC. One tag SNP in CCRN4L (rs3805213) and another tag SNP from PER3 (rs228729) demonstrated a significant correlation with genotype and allele frequency in lung cancer (P=4.4x10­3 and P=5.7x10­2; P=0.004 and P=0.02, respectively). The results of our study suggest these polymorphisms in the CCRN4L and PER3 genes may represent a risk factor in the occurrence and development of NSCLC in Brazilian patients.

Cavernostomy x resection for pulmonary aspergilloma: a 32-year history.

BACKGROUND: The most adequate surgical technique for the treatment of pulmonary aspergilloma is still controversial. This study compared two groups of patients submitted to cavernostomy and pulmonary parenchyma resection. METHODS: Cases of pulmonary aspergilloma operated upon between 1979 and 2010 were analyzed retrospectively. Group 1 consisted of patients submitted to cavernostomy and group 2 of patients submitted to pulmonary parenchyma resection. The following variables were compared between groups: gender, age, number of hospitalizations, pre- and postoperative length of hospital stay, time of follow-up, location and type of aspergilloma, preoperative symptoms, underlying disease, type of fungus, preoperative pulmonary function, postoperative complications, patient progression, and associated diseases. RESULTS: A total of 208 patients with pulmonary aspergilloma were studied (111 in group 1 and 97 in group 2). Group 1 was older than group 2. The number of hospitalizations, length of hospital stay and time of follow-up were higher in group 1. Hemoptysis was the most frequent preoperative symptom in group 1. Preoperative respiratory malfunction was more severe in group 1. Hemorrhagic complications and recurrence were more frequent in group 1 and infectious complications and residual pleural space were more common in group 2. Postoperative dyspnea was more frequent in group 2. Patient progression was similar in the two groups. No difference in the other factors was observed between groups. CONCLUSIONS: Older patients with severe preoperative respiratory malfunction and peripheral pulmonary aspergilloma should be submitted to cavernostomy. The remaining patients can be treated by pulmonary resection.

Análise da imunoexpressão da podoplanina (D2-40), da proteína nuclear Ki-67 e da catepsina D-34 e suas relações com metástase em pacientes operados por tumor carcinoide típico broncopulmonar / Analysis of the immunoexpression of podoplanin (D2-40), of the nuclear protein Ki-67, and of cathepsin D-34 and their relation to metastasis in patients operated for typical bronchopulmonary carcinoid tumor

Os tumores carcinoides broncopulmonares típicos são considerados tumores de baixo grau de malignidade, o que faz com que muitas vezes seja negligenciada sua capacidade de gerar metástases e levar a óbito. A atual impossibilidade de se fazer predições fidedignas do potencial metastático para individualizar a terapêutica e o manejo clínico do paciente portador de tumor carcinoide típico muitas vezes impõe situações de dúvida nas decisões da prática clínica diária. O conhecimento das características moleculares e genéticas desses tumores é a esperança de melhor compreensão de sua história natural, da identificação de seus fatores prognósticos e da avaliação de novas propostas terapêuticas. O objetivo deste estudo foi quantificar três imunomarcadores: o D2-40, o CD-34 e o Ki-67 e avaliar se eles são capazes de predizer metástase. Como informação adicional também foram avaliadas as variáveis clínicas e suas associações com metástases. Material e métodos: Foram analisados prontuários de 97 pacientes submetidos à cirurgia para tratamento de tumor carcinoide típico broncopulmonar e que apresentavam período de acompanhamento pós operatório de cinco anos completos. Foi estabelecido um banco de dados epidemiológicos, anátomopatológicos, cirúrgicos e clínicos relacionados à doença. Os blocos de parafina contendo os tumores primários, metastáticos e os linfonodos ressecados foram recuperados e reavaliados por dois patologistas para confirmação do diagnóstico histológico. Após confirmação diagnóstica foram realizadas as reações imunohistoquímicas para o D2-40, o CD34 e Ki-67. As variáveis demografia, gênero, idade, localização do tumor, tamanho do tumor, margem comprometida, total de linfonodos dissecados e a quantificação dos marcadores Ki-67, CD34 e podoplanina (área linfática e densidade) foram comparadas a cinco desfechos: metástase linfática; metástase em linfonodos hilares, peribrônquicos e pulmonares ipsilaterais (N1); metástase em linfonodos mediastinais...

Typical bronchopulmonary carcinoid tumors are slow-growing tumors considered to be of low malignancy, a fact that often underscores their capacity to generate metastasis that leads to death. The literature does not contain any assessment of the metastatic potential that would allow for individualized and optimized therapy that could have a positive impact upon the survival rate of patients. Genetic and biomolecular studies are the most promising venues for better comprehension of the behavior and natural history of such tumors. The objective of this investigation was to analyze three immunomarkers associated to malignity phenotypes: D2-40, CD-34, and Ki-67, and to verify whether or not they are associated to metastasis. D2-40 is a specific marker of the proliferation of the lymphatic endothelium and allows for the quantification of lymphangiogenesis. CD-34 identifies the endothelial cells of micro vessels and quantifies angiogenesis. Ki-67 in turn is a marker of neoplasia cell proliferation. As additional information several clinical variables were scrutinized for their association to metastasis. Ninety-seven subjects were assessed herein. These had undergone surgery for typical bronchopulmonary carcinoid tumor and all had complied with a full 5-year follow-up period. The paraffin blocks containing the primary and metastatic tumors and the dried lymph nodes were recovered. Once the histologic diagnosis was confirmed immunohistochemical reactions were performed for D2-40, Ki-67, and CD-34. The variables demography, gender, age, tumor site, tumor size, compromised margin, total dissected lymph nodes and quantification of D2-40 (lymphatic area and density), Ki-67 and CD-34 markers were compared to 5 outcomes: lymphatic metastasis, metastasis in hilar, peribronchic and ipsilateral pulmonary lymph nodes (N1), metastasis in ipsilateral mediastinal lymph nodes (N2), haematogenic metastasis, and lymphatic or haematogenic metastasis. It was concluded that there is...